ردیابی و شناسایی مولکولی جدایه‌های ویروئید کوتولگی رازک آلوده کننده درختان مو در برخی از مناطق ایران

نوع مقاله : مقاله پژوهشی

نویسندگان

1 بخش گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان

2 کرمان، دانشگاه شهید باهنر، دانشکده کشاورزی، بخش گیاهپزشکی

3 گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان

4 بخش گیاهپزشکی،دانشکده کشاورزی، دانشگاه شهید باهنر کرمان

چکیده

هدف: ویروئید کوتولگی رازک، (Hop stunt viroid, HSVd) گسترده‌ترین دامنه میزبانی را در بین ویروئیدها دارا می‌باشد و از مناطق مختلف دنیا گزارش شده‌است. هدف از این تحقیق، ردیابی، شناسایی و تعیین جایگاه تاکسونومیکی جدایه‌های ویروئید کوتولگی رازک در تاکستان‌های استان کرمان و برخی نقاط دیگر کشور می‌باشد.
مواد و روش‌ها: در طی تابستان سال‌های 1397 تا 1399، در مجموع 74 نمونه‌ی برگی از درختان انگور استان کرمان و دو استان آذربایجان‌شرقی (تبریز) و فارس (بیضا) جمع‌آوری شد و آلودگی نمونه‌ها به HSVd توسط آزمون RT-PCR مورد بررسی قرار گرفت. توالی ژنوم کامل هفت جدایه HSVd حاصل تعیین ترادف گردید و با توالی‌های موجود در بانک ژن مقایسه شدند. جهت بررسی روابط فیلوژنتیکی ترادف‌های بدست آمده با سایر ترادف‌های این ویروئید، از نرم افزار (version 7)MEGA  و روش neighbor-joining استفاده شد. همچنین میزان همولوژی جدایه‌ها نیز توسط نرم افزار SDT v1.2  بررسی شد. جهت تعیین دامنه میزبانی و بررسی علائم از هشت گونه و رقم گیاهی استفاده گردید.
نتایج: مطابق با گزارش‌های پیشین توالی‌های HSVd به پنج گروه فیلوژنتیکی Citrus، Hop، Plum، Plum-Hop/cit3 و Plum-Citrus تقسیم‌بندی شدند. نتایج این تحقیق نشان داد اندازه ژنوم جدایه‌های تعیین توالی شده از درختان انگور متغیر (بین 297 تا 300 نوکلئوتید) و با یکدیگر، 7/98 - 3/93 درصد تشابه داشتند. براین اساس، هفت جدایه گزارش شده در این بررسی همراه با چند جدایه گزارش شده دیگر از درختان انگور از ایران و نیز کشور آلمان در گروه اصلی Hop واقع گردیدند. سه جدایه حاصل از درختان انگور باغات استان کرمان به همراه چهار جدایه گزارش شده از این درختان، از کشور آلمان و ایران، یک زیر گروه در گروه رازک تشکیل دادند. همچنین چهار جدایه ایرانی شناسایی شده از درختان پسته در استان کرمان با تعدادی جدایه گزارش شده از درختان پسته کشور تونس (با منشاء ایرانی) نیز در گروه رازک قرار گرفتند.
نتیجه‌گیری: از آنجائی که آلودگی درختان انگور در ایران نسبت به HSVd قبل از آلودگی درختان پسته گزارش گردیده، بنابراین احتمالاً آلودگی گیاهان پسته به HSVd در ایران به‌واسطه‌ی آلودگی متقاطع و انتقال آلودگی از درختان انگور به درختان پسته رخ داده است. براین اساس با توجه به نتایج حاصل از مطالعات ملکولی و شواهد موجود، درختان انگور می‌توانند به عنوان مبداء آلودگی درختان پسته در ایران باشند.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular detection and identification of Hop stunt viroid isolates infecting grapevines in some regions of Iran

نویسندگان [English]

  • Sahar Barani 1
  • Hossein Massumi 2
  • Mohammad Maddahian 3
  • Jahangir Heydarnejad 4
  • Akbar Hosseinipour 3
1 Department of Plant Protection, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
2 Department of Plant Pathology, Shahid Bahonar University of Kerman
3 Department of Plant Pathology, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
4 : Department of plant protection, college of Agriculture, Shahid Bahonar university of Kerman
چکیده [English]

Objective
The Hop stunt viroid has the widest host range among viroids, and is prevalent worldwide. This research aims to detect and characterize HSVd variants in the vineyards of Kerman province and some other parts of Iran and analyze the variants' phylogenetic status.
Materials and methods
Seventy-four leaf samples were collected from grapevines in Kerman, East Azerbaijan (Tabriz), and Fars (Beyza) provinces during the summers of 2018–2020. The samples were processed and subjected to RT-PCR to detect HSVd using specific primers. Then, complete genomes of seven HSVd variants were sequenced and blasted into GenBank. In the MEGA 7.0 program, the neighbor-joining statistical method was used to construct a phylogenetic tree to consider relationships between the detected HSVd variants and those obtainable in GenBank. Furthermore, the homology of the grapevine-related variants was analyzed by SDT v1.2 software. Finally, eight plant species and varieties were experimentally inoculated by two HSVd variants to study symptom expression and determine host range.
Results
Throughout the world, HSVd variants have been grouped into five phylogenetic groups: Citrus, Hop, Plum, Plum-Hop/cit3, and Plum-Citrus. Accordingly, the identified seven HSVd variants in this study and several previously reported variants from grapevines in Iran and Germany were classified under the main group, Hop. The HSVd variants from grapevine had genome lengths ranging from 297 to 300 nucleotides, with 93.3–98.7% nucleotide sequence similarity. Three of the seven HSVd isolates identified in Kerman province vineyards and four isolates previously reported from grapevines in Iran and Germany formed a subgroup within the Hop cluster. In addition, the Hop group included four Iranian pistachio HSVd isolates from the Kerman province and some Tunisian pistachio HSVd isolates with Iranian provenance.
Conclusions
Given that HSVd infection of grapevines has been documented in Iran prior to pistachio infection, pistachios were most likely infected via HSVd inocula transferred from affected grapevines. Therefore, based on the molecular studies and available evidence, grapevines can be assumed to be the source of infection for pistachio trees.

کلیدواژه‌ها [English]

  • Hop stunt viroid
  • Grapevine
  • Phylogeny
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